Rumen-protected methionine during heat stress alters mTOR, insulin signaling, and 1-carbon metabolism protein abundance in liver, and whole-blood transsulfuration pathway genes in Holstein cows.

We investigated effects of rumen-protected Met (RPM) during a heat stress (HS) challenge on (1) hepatic abundance of mTOR, insulin, and antioxidant signaling proteins, (2) enzymes in 1-carbon metabolism, and (3) innate immunity. Holstein cows (n = 32; mean ± standard deviation, 184 ± 59 d in milk) were randomly assigned to 1 of 2 environmental groups, and 1 of 2 diets [total mixed ration (TMR) with RPM (Smartamine M; 0.105% dry matter as top-dress) or TMR without (CON); n = 16/diet] in a split-plot crossover design. There were 2 periods with 2 phases. During phase 1 (9 d), all cows were in thermoneutral conditions (TN; temperature-humidity index = 60 ± 3) and fed ad libitum. During phase 2 (9 d), half the cows (n = 8/diet) were exposed to HS using electric heat blankets. The other half (n = 8/diet) remained in TN, but was pair-fed to HS counterparts. After a 14-d washout and 7-d adaptation period, the study was repeated (period 2) and environmental treatments were inverted relative to phase 2, but dietary treatments were the same. Blood was collected on d 6 of each phase 2 to measure immune function and isolate whole-blood RNA. Liver biopsies were performed at the end of each period for cystathione ß-synthase (CBS) and methionine adenosyltransferase activity, glutathione concentration, and protein abundance. Data were analyzed using PROC MIXED in SAS. Abundance of CUL3, inhibitor of antioxidant responses, tended to be downregulated by HS suggesting increased oxidative stress. Heat-shock protein 70 abundance was upregulated by HS. Phosphorylated mTOR abundance was greater overall with RPM, suggesting an increase in pathway activity. An environment × diet (E × D) effect was observed for protein kinase B (AKT), whereas there was a tendency for an interaction for phosphorylated AKT. Abundance of AKT was upregulated in CON cows during HS versus TN, this was not observed in RPM cows. For phosphorylated AKT, tissue from HS cows fed CON had greater abundance compared with all other treatments. The same effect was observed for EIF2A (translation initiation) and SLC2A4 (insulin-induced glucose uptake). An E × D effect was observed for INSR due to upregulation in CON cows during HS versus TN cows fed CON or RPM. There was an E × D effect for CBS, with lower activity in RPM versus CON cows during HS. The CON cows tended to have greater CBS during HS versus TN. An E × D effect was observed for methionine adenosyltransferase, with lower activity in RPM versus CON during HS. Although activity increased in CON during HS versus TN, RPM cows tended to have greater activity during TN. Neutrophil and monocyte oxidative burst and monocyte phagocytosis decreased with HS. An (E × D) effect was observed for whole-blood mRNA abundance of CBS, SOD1 and CSAD; RPM led to upregulation during TN versus HS. Regardless of diet, CDO1, CTH, and SOD1 decreased with HS. Although HS increased hepatic HSP70 and seemed to alter antioxidant signaling, feeding RPM may help cows maintain homeostasis in mTOR, insulin signaling, and 1-carbon metabolism. Feeding RPM also may help maintain whole-blood antioxidant response during HS, which is an important aspect of innate immune function.

Residual feed intake in peripartal dairy cows is associated with differences in milk fat yield, ruminal bacteria, biopolymer hydrolyzing enzymes, and circulating biomarkers of immunometabolism.

Residual feed intake (RFI) measures feed efficiency independent of milk production level, and is typically calculated using data past peak lactation. In the current study, we retrospectively classified multiparous Holstein cows (n = 320) from 5 of our published studies into most feed-efficient (M-eff) or least feed-efficient (L-eff) groups using performance data collected during the peripartal period. Objectives were to assess differences in profiles of plasma biomarkers of immunometabolism, relative abundance of key ruminal bacteria, and activities of digestive enzymes in ruminal digesta between M-eff and L-eff cows. Individual data from cows with ad libitum access to a total mixed ration from d -28 to d +28 relative to calving were used. A linear regression model including dry matter intake (DMI), energy-corrected milk (ECM), changes in body weight (BW), and metabolic BW was used to classify cows based on RFI divergence into L-eff (n = 158) and M-eff (n = 162). Plasma collected from the coccygeal vessel at various times around parturition (L-eff = 60 cows; M-eff = 47 cows) was used for analyses of 30 biomarkers of immunometabolism. Ruminal digesta collected via esophageal tube (L-eff = 19 cows; M-eff = 29 cows) was used for DNA extraction and assessment of relative abundance (%) of 17 major bacteria using real-time PCR, as well as activity of cellulase, amylase, xylanase, and protease. The UNIVARIATE procedure of SAS 9.4 (SAS Institute Inc.) was used for analyses of RFI coefficients. The MIXED procedure of SAS was used for repeated measures analysis of performance, milk yield and composition, plasma immunometabolic biomarkers, ruminal bacteria, and enzyme activities. The M-eff cows consumed less DMI during the peripartal period compared with L-eff cows. In the larger cohort of cows, despite greater overall BW for M-eff cows especially in the prepartum (788 vs. 764 kg), no difference in body condition score was detected due to RFI or the interaction of RFI × time. Milk fat content (4.14 vs. 3.75 ± 0.06%) and milk fat yield (1.75 vs. 1.62 ± 0.04 kg) were greater in M-eff cows. Although cumulative ECM yield did not differ due to RFI (1,138 vs. 1,091 ± 21 kg), an RFI × time interaction due to greater ECM yield was found in M-eff cows. Among plasma biomarkers studied, concentrations of nonesterified fatty acids, ß-hydroxybutyrate, bilirubin, ceruloplasmin, haptoglobin, myeloperoxidase, and reactive oxygen metabolites were overall greater, and glucose, paraoxonase, and IL-6 were lower in M-eff compared with L-eff cows. Among bacteria studied, abundance of Ruminobacter amylophilus and Prevotella ruminicola were more than 2-fold greater in M-eff cows. Despite lower ruminal activity of amylase in M-eff cows in the prepartum, regardless of RFI, we observed a marked linear increase after calving in amylase, cellulase, and xylanase activities. Protease activity did not differ due to RFI, time, or RFI × time. Despite greater concentrations of biomarkers reflective of negative energy balance and inflammation, higher feed efficiency measured as RFI in peripartal dairy cows might be associated with shifts in ruminal bacteria and amylase enzyme activity. Further studies could help address such factors, including the roles of the liver and the mammary gland.

Short communication: Effect of purple corn pigment on change of anthocyanin composition and unsaturated fatty acids during milk storage.

Unsaturated fatty acids (UFA) in milk give rise to radicals and lead to lipid oxidation during storage, reducing the commercial value of milk. The objective of this study was to observe the effect of anthocyanins from purple corn pigment on the oxidation of UFA in milk. Milk samples were randomly divided into 2 groups: (1) the control (without purple corn pigment) and (2) treatment (0.3% purple corn pigment), using a completely randomized design. The milk samples were placed into plastic tubes and stored at 4°C for a period of 0, 1, 3, and 7 d. Individual anthocyanin composition and UFA were detected by HPLC-mass spectrometry and gas chromatography-mass spectrometry, respectively. The results indicated that pelargonidin (0.258 vs. 0.054 µg/mL), cyanidin (5.550 vs. 1.808 µg/mL), petunidin (0.464 vs. 0.107 µg/mL), delphinidin (2.061 vs. 0.123 µg/mL), and total anthocyanin (8.332 vs. 2.091 µg/mL) significantly decreased in response to increasing storage day. Of interest, purple corn pigment had a significant effect on most of the UFA (C14:1n-5, C16:1n-7, C18:1n-9, C18:2n-6, C18:3n-3, C18:3n-6, C20:2n-6, C20:3n-3, and C20:4n-6), except for C17:1n-7 and C20:3n-6. Specifically, various stronger positive correlations were noted for anthocyanin composition and UFA (pelargonidin and petunidin with C14:1n-5, C17:1n-7, C18:2n-6, C20:2n-6, C20:3n-3, and C20:4n-6; and cyanidin and total anthocyanins with C14:1n-5, C16:1n-7, C17:1n-7). Collectively, the current study suggested that the addition of anthocyanins from purple corn pigment had the potential to maintain UFA concentrations in milk during the storage period.

Effects of crude protein and undegradable intake protein on growth performance, nutrient utilization, and rumen fermentation in growing Thai-indigenous beef cattle.

The objective of this study was to investigate growth performance, nutrients apparent digestibility, nitrogen utilization, rumen fermentation, and rumen microorganism of growing Thai-indigenous beef cattle receiving different levels of crude protein (CP) and undegradable intake protein (UIP) diets. Eighteen healthy growing Thai-indigenous beef cattle were used in a 2 × 3 factorial randomized complete block design (RCBD). There were six treatments: two levels of CP (10% and 12% of dry matter (DM)) and three levels of UIP (15%, 25%, and 35% of CP). The results indicated that dry matter intake (DMI) and average daily gain (ADG) increased linearly (P < 0.01) with increasing UIP level. The apparent digestibility of DM, organic matter (OM), acid detergent fiber (ADF), and neutral detergent fiber (NDF); rumen fermentation parameters; microbes counts; and microbial nitrogen synthesis (MNS) were unaffected (P > 0.05) by CP and UIP levels. The nitrogen (N) intake and N retained increased linearly (P < 0.01) with increasing UIP level. Specifically, the metabolizable protein (MP) requirement of 1 g/kg BW0.75 gain was 0.34 g MP/kg BW0.75; the 10% dietary CP of DM was able to meet animals' normal nutrition requirements and 6.5% DIP of DM can provide adequate N source for the requirement of rumen microbe growth in. Collectively, it was indicated that the supplemental level of 10% CP and the ratio of UIP to DIP was 35:65 in diets shown the best growth performance for growing Thai-indigenous beef cattle under the conditions in the current study.

Short communication: Purple corn (Zea mays L.) stover silage with abundant anthocyanins transferring anthocyanin composition to the milk and increasing antioxidant status of lactating dairy goats.

The present study used 16 multiparous lactating Saanen dairy goats (body weight, 41.80 ± 2.92 kg; mean ± standard deviation) with healthy and symmetrical udders. Goats were divided into 2 blocks of 8 goats based on milk yield averaged from 75 d in milk in a randomized completed block design. The 2 study groups were the control (CSSS), in which goats were fed sticky corn stover silage, and the treatment (TPSS), in which goats were fed anthocyanin-rich purple corn (Zea mays L.) stover silage (PSS). The results indicated that the TPSS group led to an elevation in the content of milk lactose relative to the CSSS. The inclusion of anthocyanin-rich PSS had no effect on the level of 5 particular anthocyanins [i.e., cyanidin-3-glucoside, delphinidin, cyanidin, pelargonidin (Pel), as well as total anthocyanins in milk]. The pelargonidin-3-glucoside and malvidin were unable to be detected in both groups. However, the TPSS resulted in higher levels of peonidin (Peo) and malvidin-3-O-glucoside (M3G) compared with the control. Moreover, goats receiving TPSS exhibited a higher level of superoxide dismutase (SOD) in plasma and milk relative to the CSSS. Interestingly, some positive correlations were detected between the certain milk components [i.e., fat and total solids as well as fat and solids-not-fat (SNF); protein and SNF; and total solids and SNF]. In addition, the positive correlations were observed between individual anthocyanins (cyanidin-3-glucoside, delphinidin, Peo, M3G, cyanidin, and Pel) and total anthocyanins. Specifically, stronger positive correlations were noted between several antioxidant enzymes and anthocyanin composition in milk (total antioxidant capacity and Pel; SOD and Peo as well as SOD and M3G). Taken together, PSS with abundant anthocyanins can transfer anthocyanins to the milk and enhance the amount of antioxidants in lactating dairy goats.

Molecular Weight, Protein Binding Affinity and Methane Mitigation of Condensed Tannins from Mangosteen-peel (Garcinia mangostana L).

The objectives of this study were to determine the molecular weight of condensed tannins (CT) extracted from mangosteen (Garcinia mangostana L) peel, its protein binding affinity and effects on fermentation parameters including total gas, methane (CH4) and volatile fatty acids (VFA) production. The average molecular weight (Mw) of the purified CT was 2,081 Da with a protein binding affinity of 0.69 (the amount needed to bind half the maximum bovine serum albumin). In vitro gas production declined by 0.409, 0.121, and 0.311, respectively, while CH4 production decreased by 0.211, 0.353, and 0.549, respectively, with addition of 10, 20, and 30 mg CT/500 mg dry matter (DM) compared to the control (p<0.05). The effects of CT from mangosteen-peel on in vitro DM degradability (IVDMD) and in vitro N degradability was negative and linear (p<0.01). Total VFA, concentrations of acetic, propionic, butyric and isovaleric acids decreased linearly with increasing amount of CT. The aforementioned results show that protein binding affinity of CT from mangosteen-peel is lower than those reported for Leucaena forages, however, the former has stronger negative effect on IVDMD. Therefore, the use of mangosteen-peel as protein source and CH4 mitigating agent in ruminant feed requires further investigations.

Effects of supplementing rice straw with Leucaena (Leucaena leucocephala) and Madras thorn (Pithecellobium dulce) foliages on digestibility, microbial N supply and nitrogen balance of growing goats.

A total of 12 crossbred (Boer × Anglo-Nubian) goats were chosen from a commercial farm on the basis of similar body weight (25.0 ± 3.1 kg). The goats were housed in individual pens and allowed 3 weeks to adapt to experimental conditions. The goats were randomly allocated to three treatments in a 3 × 3 Latin square experiment (replicated four times). Within each period, each goat was given rice straw as roughage plus the respective treatment diet. The diets were iso-nitrogenous and iso-energetic containing cassava pulp, molasses, urea and commercial mineral and vitamin mix. The experimental treatments were (i) soybean meal (SBM), (ii) partial substitution of SBM with Leucaena (Leucaena leucocephala) foliage and (iii) partial substitution of SBM with Madras thorn (Pithecellobium dulce) foliage. Nutrient intakes, ruminal characteristics (pH, ammonia nitrogen and volatile fatty acids), nitrogen balances, plasma urea nitrogen and microbial N supply were not significantly different among treatments. The results of this study indicate that protein foliages from locally grown shrubs and trees can substitute imported feedstuff concentrates (e.g. SBM) as protein supplement for goat production.